Hydroxyl-terminated polybutadiene degrading flora and application thereof

Abstract

The invention relates to a hydroxyl-terminated polybutadiene degrading flora and application thereof, and belongs to the technical field of biology. According to the present invention, the bacterial flora capable of degrading the hydroxyl-terminated polybutadiene comprises three bacterial strains such as Agromyces sp. HA11, Microbacterium sp. HA16 and Brevibacillus sp. HA39, and the bacterial flora can be used for degrading the hydroxyl-terminated polybutadiene, the strain HA39 can effectively improve the HTPB degradation effect of agromycete HA11 and microbacterium HA16, and the improvement effect is obvious.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a hydroxyl-terminated polybutadiene-degrading bacterial community and application thereof. Background Art

[0002] Dingqiang propellant has excellent performance and high safety. It is widely used in the production of weapons and ammunition or in the preparation of aerospace engine propellants in various countries around the world. However, its storage life is limited. In addition, the rapid upgrading of weapon systems and aerospace technology in various countries in the world in recent years has inevitably produced a large amount of waste propellant. The components such as RDX, aluminum powder, and ammonium perchlorate in the waste propellant can be recycled, and the main component of the remaining bonding system is terminal hydroxyl polybutadiene (HTPB). HTPB is an artificial high molecular prepolymer with a large molecular weight and stable physical and chemical properties. If it is directly abandoned in the natural environment, it will have an adverse impact on the ecological environment and destroy the natural ecological environment.

[0003] Biodegradation is recognized as the most environmentally friendly, energy-saving and safe degradation method. At present, researchers from various countries have screened and isolated a large number of microorganisms that can degrade artificial polymer compounds from different environments such as oceans, fresh water, and soil. For example, Aspergillus niger can effectively degrade polyethylene plastics, tannins, and phytic acid; 19 species of fungi such as Candida and Fusarium can degrade petroleum hydrocarbon pollutants. A large number of studies have shown that the degrading organisms of high molecular polymers such as petroleum hydrocarbons are mostly fungi or bacteria. Among bacteria, Achromobacter, Bacillus, and Acinetobacter are the most common petroleum hydrocarbon degrading bacteria. HTPB-hydroxy-terminated polybutadiene has a structure similar to petroleum hydrocarbons, so it may be degraded by petroleum hydrocarbon degrading bacteria.

[0004] However, the effect of using a single bacterial agent to degrade HTPB is generally mediocre, and it is difficult to maintain the activity of the strain. Summary of the invention

[0005] The invention provides a bacterial community capable of efficiently degrading hydroxy-terminated polybutadiene. The bacterial community consists of three strains, namely, Agromyces sp. HA11, Microbacterium sp. HA16 and Brevibacillus sp. HA39. The Agromyces sp. HA11 and Microbacterium sp. HA16 in the bacterial community can degrade HTPB independently, and Brevibacillus sp. HA39 can break the long chain of HTPB into short chain, thereby effectively promoting the degradation of HTPB by the bacteria HA11 and HA16.

[0006] The Agromyces sp. HA11 in the present application is currently deposited in the General Microbiological Center of China National Microbiological Culture Collection Committee with a deposit number of CGMCC No. 27841 and a classification name of Agromyces sp.

[0007] The Microbacterium sp. HA16 in this application is currently strain HA16 deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration on July 10, 2023, with a deposit number of CGMCC No. 27844 and a classification name of Microbacterium sp.

[0008] The Brevibacillus sp. HA39 in this application is currently July 2023 The samples were stored at the China Microbiological Culture Collection Center on the 31st. The deposit number is CGMCC No.27842 and the classification name is Bacillus brevis.

[0009] Among them, the address of the General Microbiology Center of China Microbiological Culture Collection Administration is: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Institute of Microbiology, Chinese Academy of Sciences, Postal Code: 100101.

[0010] The strain HA39 in the present application was amplified and sequenced by PCR on the 16sDNA fragment of the strain HA39 using the first-generation sequencing technology. The sequencing results were compared with the data in the NCBI database. The comparison results showed that the similarity between the strain HA39 and Brevibacillus sp. M2 was 99.93%.

[0011] The present invention also provides a composite bacterial agent, which comprises the three strains of Agromyces sp. HA11, Microbacterium sp. HA16 and Brevibacillus sp. HA39.

[0012] The present invention also provides an application, namely, application of the above-mentioned bacterial community or the above-mentioned composite bacterial agent in preparing a product for degrading terminal hydroxyl polybutadiene.

[0013] The present invention also provides a product for degrading hydroxy-terminated polybutadiene, comprising the above-mentioned bacterial community or the above-mentioned composite bacterial agent.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention provides a bacterial community that can improve the biodegradation effect of terminal hydroxyl polybutadiene. The Agrobacterium HA11 and Microbacterium HA16 in the bacterial community can degrade HTPB independently, and the Bacillus brevis HA39 can break the long chain of HTPB into short chains, thereby effectively promoting the degradation of HTPB by the bacteria HA11 and HA16. Compared with a single strain, the bacterial community composed of three strains can significantly improve the degradation effect of hydroxyl polybutadiene. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] In order to more clearly illustrate the specific implementation methods of the present invention or the technical solutions in the prior art, the drawings required for use in the specific implementation methods or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are some implementation methods of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0017] Figure 1 is the phylogenetic tree of strain HA39 16sDNA sequence.

[0018] Figure 2 is the phylogenetic tree of the 16sDNA sequence of strain HA11.

[0019] Figure 3 is the phylogenetic tree of the 16sDNA sequence of strain HA16.

[0020] Figure 4 This is the infrared chromatogram of HTPB before degradation.

[0021] Figure 5 This is the infrared chromatogram of HTPB after degradation by the single strain HA11.

[0022] Figure 6 This is the infrared chromatogram of HTPB after degradation by the single strain HA16.

[0023] Figure 7 This is the infrared chromatogram of HTPB after degradation by the single strain HA39.

[0024] Figure 8 This is the infrared chromatogram of HTPB after degradation by bacteria (HA11, HA16 and HA39).

[0025] Fig. 9 This is the NMR spectrum of HTPB before degradation.

[0026] Fig.10 This is the nuclear magnetic resonance spectrum of HTPB after degradation by the single strain HA11.

[0027] Fig.11 This is the nuclear magnetic resonance spectrum of HTPB after degradation by the single strain HA16.

[0028] Fig.12 This is the nuclear magnetic resonance spectrum of HTPB after degradation by the single strain HA39.

[0029] Fig.13 This is the nuclear magnetic resonance spectrum of HTPB after degradation by bacterial flora (HA11, HA16 and HA39).

[0030] Fig.14This is the infrared chromatogram of HTPB after degradation by HA11 and HA16.

[0031] Specific implementation methods for screening and identifying HTPB-degrading strains

[0032] In order to make the purpose, technical scheme and beneficial effects of the present invention clearer, the screening process of the strains and the preferred embodiments of the strains in the present invention are described in detail below to facilitate the understanding of technicians.

[0033] The reagents and consumables involved in the present invention can be purchased from commercial channels. If the specific experimental conditions are not specified, they are usually carried out according to conventional experimental conditions or the conditions recommended by the reagent company.

[0034] Example 1 Isolation and screening of strain HA39

[0035] 1. Preliminary preparations for strain screening:

[0036] Prepare HTPB-inorganic salt solid culture medium, which contains only nitrogen source and no other carbon source required for microbial growth, with HTPB as the only carbon source. The specific preparation process and formula are as follows:

[0037] ①K2HPO4·3H2O 0.92g / L; ②KH2HPO4 0.7g / L; ③ZnSO4·7H2O 0.002g / L; ④

[0038] MgSO4·7H2O 0.7g / L; ⑤MnSO4 0.001g / L; ⑥NH4NO3 1.0g / L; ⑦NaCl 0.005g / L; ⑧Agar 20g / L; ①② prepared separately, sterilized at 121℃ for 30min, cooled to 50-60℃ and then mixed all reagents together.

[0039] Weigh 10 g of HTPB liquid glue, dissolve it in 50 mL of n-hexane, add 1 mL (about 0.01 g / mL) to each culture dish, and place it in a sterile operating table to air-dry for 3-4 days until the n-hexane is completely evaporated and HTPB forms a thin film on the surface of the culture medium.

[0040] 2. Screening of HTPB-degrading bacteria:

[0041] Sludge samples were collected from the sewage treatment plant, 0.01 g of sludge sample was weighed into a sterile 1.5 mL centrifuge tube, 1 mL of sterile LB culture medium was added, and the microorganisms in the sludge sample were enriched by shaking and incubating at 32°C and 140 rpm / min for 24 h. The sludge sample microorganism enrichment culture medium was diluted to 10 -4, 100 μL of each dilution was applied to the prepared HTPB-inorganic salt solid culture medium, and cultured in a 32°C constant temperature incubator for 30 days to screen HTPB-degrading bacteria.

[0042] 3. Purification of HTPB-degrading bacteria:

[0043] LB solid medium was prepared to streak-purify the colonies of strain HA39. After purification for 3-4 times, a pure strain was finally obtained and preserved, and named HA39.

[0044] Example 2 Identification of strain HA39

[0045] 1. Morphological identification:

[0046] The screened HA39 strain grew fast in LB solid culture dishes, had large colonies, was colorless, had a smooth surface, and had irregular edges.

[0047] 2. Molecular biological identification of strains:

[0048] The DNA of HA39 strain was extracted, PCR amplified and the amplified products were sequenced.

[0049] Primer: 27F (5′-AGAGTTTGATCCTGGCTCAG-3′;

[0050] 1492R(5'-TACGACTTAACCCCAATCGC-3'.

[0051] Reaction system: 2×TaqPCR Master Mix 12.5μL, upstream and downstream primers 1μL each, DNA template 1μL, ddH2O10μL; reaction program: 95℃5min; 94℃1min, 55℃1min, 72℃90s, 35 cycles; 72℃10min.

[0052] The PCR products were sent to Shanghai Bioengineering Co., Ltd. for sequencing, and the sequencing data were spliced ​​and compared with NCBI to identify the species of the degrading strain.

[0053] The sequencing and assembly results of strain HA39 16sDNA are shown in SEQ ID NO.1.

[0054] The 16sDNA sequencing results of strain HA39 showed that it was Brevibacillus sp. M2. A phylogenetic tree was constructed using 16sDNA sequencing data. The results are as follows: Figure 1As shown, the results showed that the HTPB-degrading strain HA39 was most closely related to the Brevibacillus agri strain M1-5. Brevibacillus borstelensis strain: DSM 6347T, Brevibacillus fulvus strain: K282 and other Brevibacillus strains were closely related to the strain HA39 and were therefore clustered in the same large branch. There was a certain degree of genetic relationship between Ammonii agariperforans, Fontibacillus sp. enrichment culture clone DF60, and Cohnella sp. T (genus) and the strain HA39, and Clostridium bacteria were distantly related to HA39.

[0055] Example 3 Isolation and screening of strain HA11

[0056] 1. Preliminary preparations for strain screening:

[0057] Prepare HTPB-inorganic salt solid culture medium, which contains only nitrogen source and no other carbon source required for microbial growth, with HTPB as the only carbon source. The specific preparation process and formula are as follows:

[0058] ①K2HPO 4. 3H2O 0.92g / L; ②KH2HPO4 0.7g / L; ③ZnSO4.7H2O 0.002g / L; ④MgSO 4. 7H2O0.7g / L; ⑤MnSO4 0.001g / L; ⑥NH4NO3 1.0g / L; ⑦NaCl 0.005g / L; ⑧Agar 20g / L; ①② prepared separately, sterilized at 121℃ for 30min, cooled to 50-60℃ and then mixed all reagents together.

[0059] Weigh 10 g of HTPB liquid glue, dissolve it in 50 mL of n-hexane, add 1 mL (about 0.01 g / mL) to each culture dish, and place it in a sterile operating table to air-dry for 3-4 days until the n-hexane is completely evaporated and HTPB forms a thin film on the surface of the culture medium.

[0060] 2. Screening of HTPB-degrading bacteria:

[0061] Sludge samples were collected from the sewage treatment plant, 0.01 g of sludge sample was weighed into a sterile 1.5 mL centrifuge tube, 1 mL of sterile LB culture medium was added, and the microorganisms in the sludge sample were enriched by shaking and incubating at 32°C and 140 rpm / min for 24 h. The sludge sample microorganism enrichment culture medium was diluted to 10 -4 , 100 μL of each dilution was applied to the prepared HTPB-inorganic salt solid culture medium, and cultured in a 32°C constant temperature incubator for 30 days to screen HTPB-degrading bacteria.

[0062] 3. Purification of HTPB-degrading bacteria:

[0063] LB solid medium was prepared to streak-purify the colonies of strain HA11. After purification for 3-4 times, a pure strain was finally obtained and preserved, and named HA11.

[0064] Example 4 Identification of strain HA11

[0065] 1. Morphological identification:

[0066] like Figure 2 As shown, the screened HA11 strain has a fast growth rate, large colonies, light yellow color, smooth surface and neat edges in LB solid culture dishes.

[0067] 2. Molecular biological identification of strains:

[0068] The DNA of HA11 strain was extracted, PCR amplified and the amplified products were sequenced.

[0069] Primer: 27F (5′-AGAGTTTGATCCTGGCTCAG-3′;

[0070] 1492R(5'-TACGACTTAACCCCAATCGC-3'.

[0071] Reaction system: 2×TaqPCR Master Mix 12.5μL, upstream and downstream primers 1μL each, DNA template 1μL, ddH2O10μL; reaction program: 95℃5min; 94℃1min, 55℃1min, 72℃90s, 35 cycles; 72℃10min.

[0072] The PCR products were sent to Shanghai Bioengineering Co., Ltd. for sequencing, and the sequencing data were spliced ​​and compared with NCBI to identify the species of the degrading strain.

[0073] The results of 16sDNA sequencing and assembly of strain HA11 are shown in SEQ ID NO.2;

[0074] The 16sDNA fragment of strain HA11 was amplified by PCR and sequenced by the first generation sequencing technology. The sequencing results were compared with the data in the NCBI database. The comparison results showed that the similarity between strain HA11 and Agromyces p. 3098BRRJ was 99.21%. Figure 2 The phylogenetic tree was constructed using 16sDNA sequencing data, and the results showed that the HTPB-degrading strain HA11 was closely related to Agromyces sp. strain UASWS3226 AD1SH4, Agromyces agglutinans strain CFH 90414, and Agromyces agglutinans strain CFH

[0075] 90414) and other strains are closely related and therefore clustered in the same large branch; mnibacterium sp. VN08A0400, Crocebacterium sp. SS6.7, and Diaminobutyricimonas massiliensis strain FF2 are distantly related species of strain HA11.

[0076] Deposit information: The strain HA11 is currently deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, address: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101, with the deposit number CGMCC No.27841 and the classification name of Agrobacterium spp.

[0077] Example 5 Isolation and screening of strain HA16

[0078] 1. Preliminary preparations for strain screening:

[0079] Prepare HTPB-inorganic salt solid culture medium, which contains only nitrogen source and no other carbon source required for microbial growth, with HTPB as the only carbon source. The specific preparation process and formula are as follows:

[0080] ①K2HPO4·3H2O 0.92g / L; ②KH2HPO4 0.7g / L; ③ZnSO4·7H2O 0.002g / L; ④MgSO4·

[0081] 7H2O 0.7g / L; ⑤MnSO4 0.001g / L; ⑥NH4NO3 1.0g / L; ⑦NaCl 0.005g / L; ⑧Agar 20g / L; ①② prepared separately, sterilized at 121℃ for 30min, cooled to 50-60℃ and then mixed all reagents together.

[0082] Weigh 10 g of HTPB liquid glue, dissolve it in 50 mL of n-hexane, add 1 mL (about 0.01 g / mL) to each culture dish, and place it in a sterile operating table to air-dry for 3-4 days until the n-hexane is completely evaporated and HTPB forms a thin film on the surface of the culture medium.

[0083] 2. Screening of HTPB-degrading bacteria:

[0084] Sludge samples were collected from the sewage treatment plant, 0.01 g of sludge sample was weighed into a sterile 1.5 mL centrifuge tube, 1 mL of sterile LB culture medium was added, and the microorganisms in the sludge sample were enriched by shaking and incubating at 32°C and 140 rpm / min for 24 h. The sludge sample microorganism enrichment culture medium was diluted to 10 -4 , 100 μL of each dilution was applied to the prepared HTPB-inorganic salt solid culture medium, and cultured in a 32°C constant temperature incubator for 30 days to screen HTPB-degrading bacteria.

[0085] 3. Purification of HTPB-degrading bacteria:

[0086] LB solid medium was prepared to streak and purify the colonies of strain HA16. After purification for 3-4 times, a pure strain was finally obtained and preserved, and named HA16.

[0087] Example 6 Identification of strain HA16

[0088] 1. Morphological identification:

[0089] The screened HA16 strain grew fast in the LB solid culture dish, had large colonies, was light yellow, had a smooth surface and neat edges.

[0090] 2. Molecular biological identification of strains:

[0091] The DNA of HA16 strain was extracted, PCR amplified and the amplified products were sequenced.

[0092] Primer: 27F (5′-AGAGTTTGATCCTGGCTCAG-3′;

[0093] 1492R(5'-TACGACTTAACCCCAATCGC-3'.

[0094] Reaction system: 2×TaqPCR Master Mix 12.5μL, upstream and downstream primers 1μL each, DNA template 1μL, ddH2O10μL; reaction program: 95℃5min; 94℃1min, 55℃1min, 72℃90s, 35 cycles; 72℃10min.

[0095] The PCR products were sent to Shanghai Bioengineering Co., Ltd. for sequencing, and the sequencing data were spliced ​​and compared with NCBI to identify the species of the degrading strain.

[0096] The sequencing and assembly results of strain HA16 16sDNA are shown in SEQ ID NO.3.

[0097] The 16sDNA fragment of strain HA16 was amplified by PCR and sequenced by the first generation sequencing technology. The sequencing results were compared with the data in the NCBI database. The comparison results showed that the similarity between strain HA16 and Microbacterium sp. HBUM179221 was 99.21%. Figure 3 The phylogenetic tree was constructed using 16sDNA sequencing data, and the results showed that the HTPB-degrading strain HA16 was closely related to strains such as Clavibactermichiganensis michiganensis LMG 7333 and Microbacteriaceae Agrococcus Jejuensis strain SSW1-48, so they were clustered in the same large branch; Cellulomonas sp. IFO16243, Frankia sp., and Thermolithobacter ferrireducens strain JW / KA-2 were distant sources of strain HA16.

[0098] Deposit information: The strain HA16 was currently deposited at the General Microbiology Center of the China Culture Collection Administration on July 10, 2023, at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing, Postal Code: 100101, with the deposit number CGMCC No. 27844 and the classification name Microbacterium.

[0099] Example 7 Testing of the effect of strain HA39 on improving the degradation of HTPB by strains HA11 and HA16

[0100] Example 1:

[0101] 1. Prepare LB liquid culture medium, the composition ratio of which is: Tryptone 10 g / L, yeast extract 5 g / L, sodium chloride (NaCl) 10 g / L, pH 7.0-7.4.

[0102] 2. Prepare sterile HTPB-inorganic salt liquid culture medium with the following formula: K2HPO4·3H2O 0.92g / L, KH2HPO4 0.7g / L, FeSO4·7H2O 0.002g / L, MgSO4·7H2O 0.7g / L, MnSO4 0.001g / L, NH4NO3 1.0g / L, NaCl 0.005g / L, ZnSO4·7H2O 0.002g / L. Prepare 150mL conical flasks, spread 0.1g HTPB liquid gel in each flask, sterilize at 120℃ for 30min, and then add 100mL sterile HTPB-inorganic salt liquid culture medium in a sterile environment.

[0103] The purified strain HA39 single colony obtained in Example 1 and the strains HA11 and HA16 were picked and inoculated into LB liquid medium for culture, and cultured at 32°C and 140 rpm / min for 24 h until the OD 600 When the pH value is ≈1.0, add 1 mL of the bacterial suspension of strains HA11, HA16, and HA39 to 100 mL of inorganic salt liquid culture medium containing 0.1 g HTPB in a ratio of 1:1:1. Seal the bottle mouth with sealing film and tin foil. Culture at 32°C and 140 rpm / min for 28 days. Extract the fermentation liquid with n-hexane ether for 3 times, then extract it with chloroform for 3 times, and place it in a ventilated medium until the n-hexane and chloroform are completely evaporated. All the extraction products are placed in the same sample bottle.

[0104] Comparative Example 1:

[0105] The difference from Example 1 is that HA39 bacterial solution was not inoculated into the HTPB inorganic salt medium, but 3 mL of sterile water was added.

[0106] Comparative Example 2:

[0107] The difference from Example 1 is that only 3 ml of HA11 bacterial solution is inoculated into the HTPB inorganic salt medium.

[0108] Comparative Example 3:

[0109] The difference from Example 1 is that only 3 ml of HA16 bacterial solution is inoculated into the HTPB inorganic salt medium.

[0110] Comparative Example 4:

[0111] The difference from Example 1 is that only 1.5 ml of HA16 bacterial solution and 1.5 ml of HA11 bacterial solution were inoculated into the HTPB inorganic salt medium.

[0112] The experimental results are as follows:

[0113] like Figure 4-8 As shown, the fermentation broth after degradation of single strains (strain HA11 and strain HA16) and composite bacterial consortium (HA11, HA16 and HA39; HA11 and HA16) was extracted with n-hexane and chloroform, respectively, and the HTPB degradation products were recovered. After the extraction solvents (chloroform and n-hexane) were evaporated, infrared chromatography analysis was performed. Figure 4 The infrared spectrum of HTPB not treated with degradation strains showed no obvious changes.

[0114] like Figure 5-7 As shown in the figure, in addition to the fact that the HTPB degraded by the fungus HA11 can oxygenate the long chain of HTPB during the degradation process, catalyzing the generation of a small amount of free hydroxyl groups ( Figure 5 :384.16-3681.90cm -1 ), the remaining strains did not add oxygen to produce free hydroxyl groups when degrading HTPB alone.

[0115] Fig.14 The infrared chromatogram showed that the HTPB changed little after being treated with the two strains of strain HA11 and strain HA16; further, Figure 4-Figure 8 The infrared chromatogram shows that the infrared spectrum of HTPB after degradation by the composite bacteria (HA11, HA16 and HA39) changes significantly, producing a large amount of alcohol and phenolic substances (about 3300-3500cm -1 ).

[0116] Fig. 9 This is the NMR spectrum of HTPB without degradation strain treatment. Fig.10 The nuclear magnetic resonance spectrum showed that strain HA11 would slightly interrupt the long-chain structure of HTPB during the degradation of HTPB, so some -CH3 structures could be detected at 1.5ppm; the signal intensity was about 5.01E+07;

[0117] Fig.11 It shows that strain HA16 also breaks the long chain structure of HTPB during the HTPB degradation process. The 1.5ppm signal is enhanced, and the -CH3 structure is detected with a signal intensity of 3.2E+08. However, its efficiency in breaking the long chain of HTPB is slightly lower than that of strain HA39. The peak values ​​of HTPB treated with strain HA39 also change at 5.5ppm-5.0ppm and 1.5ppm-1.0ppm ( Fig.12), the peak of HTPB at 5.5ppm after being treated with strain HA39 was significantly weakened, indicating that the strain destroyed the original aldehyde structure in the process of growing with HTPB; in addition, during the degradation process, strain HA39 destroyed the original long-chain structure of HTPB and degraded HTPB into a large number of short-chain structures, thus producing a large amount of -CH3. Therefore, the peak signal at 1.5ppm of the HTPB nuclear magnetic spectrum after being treated with strain HA39 was significantly enhanced by about 4.3E+08, which was stronger than that of strains HA11 and HA16;

[0118] The nuclear magnetic resonance spectra showed that the signal intensity of -CH3 detected in HTPB treated with strain HA11 and strain HA16 was about 2.9E+08.

[0119] Fig.13 It is HTPB degraded by a composite bacterial community (HA11, HA39, HA16). The signal intensity of -CH3 after degradation by three strains is about 5.5E+07.

[0120] The comprehensive test results (NMR and IR) showed that strains HA16 and HA11 could both utilize HTPB with a destroyed long-chain structure for growth. However, although strain HA11 could produce a variety of degradation products in the process of degrading HTPB, the efficiency of this strain in breaking the long chain of HTPB was low. When HA16 treated HTPB alone, it would mainly focus on breaking the long-chain structure of HTPB, but the strain mainly broke HTPB into medium-long chains, and only a small amount of broken HTPB was used by strain HA16; strain HA39 could break the long chain of HTPB into a large number of short-chain structures, but it itself rarely used short-chain HTPB for growth. Adding the degrading bacteria of strain HA39 can effectively destroy the long-chain and medium-long chain structures of HTPB, which is beneficial for strains HA11 and HA16 to degrade HTPB, and further improve the degradation effect of HTPB.

[0121] Finally, it should be noted that the above preferred embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail through the above preferred embodiments, those skilled in the art should understand that various changes can be made in form and details without departing from the scope defined by the claims of the present invention.

Claims

1. A bacterial community capable of degrading hydroxy-terminated polybutadiene, characterized in that: The invention is composed of three strains, namely Agromyces sp. HA11, Microbacterium sp. HA16 and Brevibacillus sp. HA39; all strains are deposited in China General Microbiological Culture Collection Center; the deposit number of Agromyces sp. HA11 is CGMCC No. 27841, and its classification name is Agromyces; the deposit number of Microbacterium sp. HA16 is CGMCC No. 27844, and its classification name is Microbacterium; the deposit number of Brevibacillus sp. HA39 is CGMCC No. 27842, and its classification name is Brevibacillus sp.

2. The bacterial flora capable of degrading hydroxy-terminated polybutadiene according to claim 1, characterized in that: The effective bacterial count ratio of the three strains of Agrobacterium HA11, Microbacterium HA16 and Bacillus brevis HA39 is 1:1:

1.

3. A composite bacterial agent, characterized in that: Comprising the bacterial flora according to claim 1.

4. A composite bacterial agent according to claim 3, characterized in that: The composite bacterial agent is obtained by mixing corresponding strains in a ratio of 1:1:

1.

5. Use of the bacterial community according to claim 1 or the composite bacterial agent according to claim 3 in preparing a product for degrading hydroxy-terminated polybutadiene.

6. A product for degrading hydroxy-terminated polybutadiene, characterized in that: It includes the bacterial community described in claim 1 or the composite bacterial agent described in claim 3.

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